Multiplexed on-chip real-time PCR using hydrogel spot array for microRNA profiling of minimal tissue samples

Jung

Adv Healthcare Materials

et al. 2020

Self Cite

Rapid and simple detection of RNA targets is in high demand due to the growing threat of pandemic viruses. One‐step real‐time, reverse transcription–polymerase chain reaction (One‐step RT‐qPCR) using a controlled release system of thermo‐responsive materials is developed in this paper to enable high‐fidelity RNA analysis as suppressing by‐products. The nanocapsules, consisting of upper critical solution temperature (UCST) material and PCR primers, carry or release the primers depending upon the temperature. The UCST nanocapsules are introduced into hydrogel microparticles incorporated with RT primers and then the target RNA is selectively amplified in the microparticle through one‐step RT‐qPCR. Severe side products are sharply subdued by separating the PCR primers from the RT process by means of the microparticles with nanocapsules. Because the one‐step assay is now implemented in a single microparticle, multiple target RNAs can be analyzed in a simple RT‐qPCR of multiple particles. Reliable 18‐plex one‐step RT‐qPCR is successfully conducted within 30 min using single‐color fluorescent optics. This work also explains the facile fabrication processes used for the thermo‐responsive nanocapsules and hydrogel microparticles by the blending polymerization method. Extensible multiplex analysis of influenza virus demonstrates the versatile uses of this one‐step RT‐qPCR platform.

Section: Figurementioning

confidence: 99%

Section: Figurementioning

confidence: 99%

Thermo‐Responsive Polymer Capsules in Real‐Time One‐Step RT‐PCR for Highly Multiplex RNA Analysis

Jung

Adv Healthcare Materials

et al. 2020

Self Cite

“…Over the last several decades, there have been numerous studies that have found associations between miRNAs and various types of cancer, such as lymphocytic leukemia [4,5], lung cancer [6,7], colorectal neoplasia [8], Burkitt lymphoma [9], glioblastoma [10], tumor cell [11], B cell lymphomas of hepatocellular carcinoma (HCC) [12], oral squamous cell carcinoma [13], and breast cancer [14]. Relatedly, many biological techniques have been developed for miRNA detectors, including Northern blot [15], real-time PCR [16], oligonucleotide microarray [17,18], laser-induced fluorescence [19], microarray [20,21], and even various label-free techniques [22], as well as some strategy approaches for the detection of miRNAs, such as electrochemical, photoelectrochemical and optical fiber sensing methods. The purpose of this study is to review the literature on photoelectrochemical biosensors for miRNA assay that show potential to be applied in bioanalysis research.…”

Section: Introductionmentioning

confidence: 99%

A Lamping U-Shaped Fiber Biosensor Detector for MicroRNA

Wen¹,

Huang²,

Li³

et al. 2020

Sensors

This study presents a U-shaped optical fiber developed for a facile application of microRNA detection. It is fabricated by the lamping process and packaged in a quartz tube to eliminate human negligence. In addition, silanization and electrostatic self-assembly are employed to bind gold nanoparticles and miRNA-133a probe onto the silicon dioxide of the fiber surface. For Mahlavu of hepatocellular carcinoma (HCC), detection is determined by the wavelength shift and transmission loss of a U-shaped optical fiber biosensor. The spectral sensitivity of wavelength and their coefficient of determination are found at −218.319 nm/ ng/mL and 0.839, respectively. Concurrently, the sensitivity of transmission loss and their coefficient of determination are found at 162.394 dB/ ng/mL and 0.984, respectively. A method for estimating the limit of detection of Mahlavu is at 0.0133 ng/mL. The results show that the proposed U-shaped biosensor is highly specific to miRNA-133a and possesses good sensitivity to variations in specimen concentration. As such, it could be of substantial value in microRNA detection.

“…Several methods have been generally applied for the quantification of microRNAs, such as northern blot [9], real-time polymerase chain reaction [10] and microarrays [11]. Nevertheless, they present some disadvantages, including high cost, time-consuming, requirement of a qualified personnel and sophisticated instrumentation.…”

Section: Introductionmentioning

confidence: 99%

A Highly Sensitive Electrochemical Biosensor Based on Carbon Black and Gold Nanoparticles Modified Pencil Graphite Electrode for microRNA-21 Detection

Yammouri

Mohammadi

2019

Chemistry Africa

This work aims to develop cost-effective, simple and sensitive electrochemical biosensor for detection of microRNA-21. The combination of carbon black (CB) and gold nanoparticles (AuNPs) as nanohybrid was employed for the first time as a platform for microRNA-21 biosensor fabrication. The developed biosensor is based on the immobilization of thiolated capture probe (complementary sequence of microRNA-21) labeled with methylene blue on the surface of the pencil graphite electrode modified with CB/AuNPs nanohybrid. After hybridization with the target microRNA-21, the orientation of the labeled capture probe changed which causes a decrease of the response of methylene blue oxidation. Differential pulse voltammetry was used for monitoring the methylene blue response before and after hybridization. Under the optimal conditions, the developed biosensor was characterized using differential pulse voltammetry (DPV), cyclic voltammetry (CV) and electrochemical impedance spectroscopy (EIS). The detection of microRNA-21 was carried out using a DPV. A wide linear range was obtained between 2.9 fM and 0.7 µM of microRNA-21. The calculated limit of detection was 1 fM of microRNA-21. This approach shows a good reproducibility, stability and an excellent selectivity. The proposed biosensor was used for microRNA-21 analysis in serum and a satisfactory result was obtained.